A new method for phenotyping proliferating cell nuclear antigen positive cells using flow cytometry: implications for analysis of the immune response in vivo

Kühn, Ulrich; Lempertz, Uwe; Knop, Jürgen; Becker, Detlef

doi:10.1016/0022-1759(94)00287-7

Cited by 18 publications

(7 citation statements)

References 12 publications

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“…This can be useful in identifying DNA synthesis inhibition effects and cellcycle related anomalies known to be induced by certain drugs [10], but otherwise not studied, especially in allergy. Nevertheless, when no such interferences exist, PCNA expression has been shown to correlate very well with thymidine incorporation [11]and this was confirmed in our own preliminary experiments (data not shown). Another advantage of PCNA is the opportunity for simultaneous evaluation of the immunophenotype of proliferating cells.…”

Section: Discussionsupporting

confidence: 70%

Correlation of Lymphocyte Proliferating Cell Nuclear Antigen Expression with Dietary Cow’s Milk Antigen Load in Infants with Allergy to Cow’s Milk

Papadopoulos

Syrigou

Bossios

et al. 1999

Int Arch Allergy Immunol

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Background: Controversial results have been reported on the participation and diagnostic value of lymphocyte reactivity in cow’s milk (CM) allergy. In this study, we used a specific nuclear marker to evaluate lymphocyte proliferation in IgE–mediated CM allergy in infants, and examine its relation with diets containing different CM antigen loads. Methods: Infants with IgE–mediated CM allergy, as assessed by open provocation and RAST, were grouped according to their exclusive diet, either CM formulae, breast feeding, or hydrolysed whey formulae. A group of non–atopic infants receiving CM was also examined. Lymphocyte proliferation to β–lactoglobulin was evaluated by quantitation of the proliferating cell nuclear antigen (PCNA) expression in peripheral blood mononuclear cells, by flow cytometry. Immunophenotypic surface markers were also examined. Results: A marked difference of PCNA expression between CM–fed allergic infants and healthy controls was observed (p<0.001). In this setting, PCNA expression ≥10% was highly specific and sensitive as a marker of CM allergy in CM–fed infants. Moreover, a significant correlation (p<0.001) between antigen load and PCNA was established in CM–allergic infants under different diets, higher values obtained with increasing antigen loads. In addition, within the group fed hydrolyzed formulae, low–molecular–weight products resulted in marginally lower PCNA expression than higher–molecular–weight formulae. No differences in immunophenotype were found, with the exception of a higher CD23 expression in the breast–fed group. Conclusions: PCNA could be a useful marker in the assessment of lymphocyte proliferation to CM antigens. Low CM antigen diets are related with reduced lymphocyte reactivity, which may partly explain the clinical benefit observed with such diets.

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Section: Discussionsupporting

confidence: 70%

Correlation of Lymphocyte Proliferating Cell Nuclear Antigen Expression with Dietary Cow’s Milk Antigen Load in Infants with Allergy to Cow’s Milk

Papadopoulos

Syrigou

Bossios

et al. 1999

Int Arch Allergy Immunol

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“…Kuhn et al (1995) described the good correlation between the incorporation of 3H-TdR and the nuclear expression of PCNA in LLNA. They stated that the total population of PCNA positive cells was significantly increased after application of contact sensitizers (DNFB, TNCB, oxazolone) in comparison to solvent-treated mice (Kuhn et al, 1995). In addition, they stained LNC of mice treated with oxazolone for PCNA and the T cell markers CD4 and CD8.…”

Section: Discussionmentioning

confidence: 99%

“…Boussiquet-Levoux et al (1995) reported that a proliferating response of lymph node cells (LNC) in mice was assessed by immunohistochemistry, using bromodeoxyuridine (BrdU). The proliferation of LNC was also determined by the flow cytometric (FCM) analysis of proliferating cell nuclear antigen (PCNA) (Kuhn et al, 1995). Other than proliferating responses, several functional approaches were reported, including phenotypic analysis of LNC subpopulations: B220 positive cells (Sikorski et al, 1996), CD3 positive cells and CD25 positive cells (De Silva et al, 1993), CD62L lo /CD44 hi double positive cells (Gerberick et al, 1997(Gerberick et al, , 1999a, CD86 positive cells, and I-A k positive cells (Gerberick et al, 1999b).…”

Section: Introductionmentioning

confidence: 99%

Local lymph node assay with non-radioisotope alternative endpoints.

et al. 2002

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-The local lymph node assay has recently been accepted by regulatory agencies as a standalone alternate method for predicting allergic contact dermatitis. To compare the sensitivity of non-radioisotope methods with that of the standard assay, we determined if these modified methods would affect evaluation of sensitization potency. For this reason, we used 2,4-dinitrochlorobenzene (DNCB) and benzocaine for different sensitizing criteria.Female CBA mice were treated for 3 days with a test compound or vehicle applied to each side of both ears. Bilateral auricular lymph node proliferative activity was assessed by the following endpoints with incorporation of 3 H-methyl thymidine (3H-TdR), bromodeoxyuridine (BrdU) in vivo, and BrdU ex vivo, IL-2 production, and proliferating cell nuclear antigen (PCNA) expression. Ear thickness was also tested.The strong sensitizer DNCB was detectable by any of the non-radioisotope endpoints as well as by radioisotope-dependent standard assay. On the other hand, when evaluating the weak sensitizer benzocaine, significant changes were evident in BrdU incorporation ex vivo and in vivo, and IL-2 production. We believe that these non-radioisotope methods can assess allergic contact dermatitis caused by chemicals even in the laboratory, where it can be difficult to handle radioisotopes.

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“…Flow cytometric analysis was used in order to measure lymphocyte proliferation. We measured expression of proliferating cell nuclear antigen (PCNA), an auxiliary cyclin protein necessary for DNA polymerase activity, maximally expressed in mid S-phase as previously proposed by Kühn et al (1995). Proliferative activity was evaluated after 72 h of incubation, using anti-PCNA monoclonal antibody.…”

Section: Proliferation Assaymentioning

confidence: 99%

Melatonin protects rat thymus against oxidative stress caused by exposure to microwaves and modulates proliferation/apoptosis of thymocytes

Sokolović

Djordjević²,

Kocić³

et al. 2013

gpb

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The aim of the study was to evaluate the effect of melatonin on oxidative stress, DNA fragmentation, apoptsis and proliferation in thymus tissue of rats exposed to microwaves. Wistar rats were divided in four groups: I - treated with saline; II - treated with melatonin; III - microwaves exposed; IV - microwaves exposed and melatonin treated. Melatonin (2 mg/kg i.p.) was administered daily. Animals were sacrificed after 20, 40 and 60 days. A significant increase in malondialdehyde and carbonyl group content, as well as decrease in catalase and increase in xanthine oxidase activity were registered under microwave exposure. Melatonin prevented the increase in malondialdehyde and carbonyl group content, and reversed the effect on catalase and xanthine oxidase activity. Both, alkaline and acid DNase activity were increased due to microwave exposure. Furthermore, microwaves caused increase in apoptosis rate (detected using Annexin V-FITC/PI kit) and reduced proliferative capacity of thymocytes (induced by ConA). However, melatonin caused decrease in alkaline and acid DNase activity, decrease in apoptotic rate and increase in proliferation rate of thymocytes. Melatonin exerts protective effects on rat thymocytes by modulating processes of apoptosis and proliferation, and causes decrease in DNA fragmentation and oxidative stress intensity under exposure to microwaves.

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A new method for phenotyping proliferating cell nuclear antigen positive cells using flow cytometry: implications for analysis of the immune response in vivo

Cited by 18 publications

References 12 publications

Correlation of Lymphocyte Proliferating Cell Nuclear Antigen Expression with Dietary Cow’s Milk Antigen Load in Infants with Allergy to Cow’s Milk

Correlation of Lymphocyte Proliferating Cell Nuclear Antigen Expression with Dietary Cow’s Milk Antigen Load in Infants with Allergy to Cow’s Milk

Local lymph node assay with non-radioisotope alternative endpoints.

Melatonin protects rat thymus against oxidative stress caused by exposure to microwaves and modulates proliferation/apoptosis of thymocytes

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A new method for phenotyping proliferating cell nuclear antigen positive cells using flow cytometry: implications for analysis of the immune response in vivo

Cited by 18 publications

References 12 publications

Correlation of Lymphocyte Proliferating Cell Nuclear Antigen Expression with Dietary Cow’s Milk Antigen Load in Infants with Allergy to Cow’s Milk

Correlation of Lymphocyte Proliferating Cell Nuclear Antigen Expression with Dietary Cow’s Milk Antigen Load in Infants with Allergy to Cow’s Milk

Local lymph node assay with non-radioisotope alternative endpoints.

Melatonin protects rat thymus against oxidative stress caused by exposure to microwaves and modulates proliferation/apoptosis of thymocytes

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Product

Resources

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Melatonin protects rat thymus against oxidative stress caused by exposure to microwaves and modulates proliferation/apoptosis of thymocytes